Microfluidic immunosensor for rapid and highly-sensitive salivary cortisol quantification.

This paper presents a novel poly(dimethylsiloxane) (PDMS) microfluidic immunosensor that integrates a complementary metal-oxide-semiconductor (CMOS) optical detection system for a rapid and highly-sensitive quantification of salivary cortisol. The simple and non-invasive method of saliva sampling provides an interesting alternative to the blood, allowing a fast sampling at short intervals, relevant for many clinical diagnostic applications. The developed approach is based on the covalent immobilization of a coating antibody (Ab), a polyclonal anti-IgG, onto a treated PDMS surface. The coating Ab binds the capture Ab, an IgG specific for cortisol, allowing its correct orientation. Horseradish peroxidase (HRP)-labelled cortisol is added to compete with the cortisol in the sample, for the capture Ab binding sites. The HRP-labelled cortisol, bonded to the capture Ab, is measured through the HRP enzyme and the tetramethylbenzidine (TMB) substrate reaction. The cortisol quantification is performed by colorimetric detection of HRP-labelled cortisol, through optical absorption at 450nm, using a CMOS silicon photodiode as the photodetector. Under the developed optimized conditions presented here, e.g., microfluidic channels geometry, immobilization method and immunoassay conditions, the immunosensor shows a linear range of detection between 0.01-20ng/mL, a limit of detection (LOD) of 18pg/mL and an analysis time of 35min, featuring a great potential for point-of-care applications requiring continuous monitoring of the salivary cortisol levels during a circadian cycle.

Evaluation of the successive approximations method for acoustic streaming numerical simulations.

This work evaluates the successive approximations method commonly used to predict acoustic streaming by comparing it with a direct method. The successive approximations method solves both the acoustic wave propagation and acoustic streaming by solving the first and second order Navier-Stokes equations, ignoring the first order convective effects. This method was applied to acoustic streaming in a 2D domain and the results were compared with results from the direct simulation of the Navier-Stokes equations. The velocity results showed qualitative agreement between both methods, which indicates that the successive approximations method can describe the formation of flows with recirculation. However, a large quantitative deviation was observed between the two methods. Further analysis showed that the successive approximation method solution is sensitive to the initial flow field. The direct method showed that the instantaneous flow field changes significantly due to reflections and wave interference. It was also found that convective effects contribute significantly to the wave propagation pattern. These effects must be taken into account when solving the acoustic streaming problems, since it affects the global flow. By adequately calculating the initial condition for first order step, the acoustic streaming prediction by the successive approximations method can be improved significantly.

Improving acoustic streaming effects in fluidic systems by matching SU-8 and polydimethylsiloxane layers.

This paper reports the use of acoustic waves for promoting and improving streaming in tridimensional polymethylmethacrylate (PMMA) cuvettes of 15mm width×14mm height×2.5mm thickness. The acoustic waves are generated by a 28µm thick poly(vinylidene fluoride) - PVDF - piezoelectric transducer in its ß phase, actuated at its resonance frequency: 40MHz. The acoustic transmission properties of two materials - SU-8 and polydimethylsiloxane (PDMS) - were numerically compared. It was concluded that PDMS inhibits, while SU-8 allows, the transmission of the acoustic waves to the propagation medium. Therefore, by simulating the acoustic transmission properties of different materials, it is possible to preview the acoustic behavior in the fluidic system, which allows the optimization of the best layout design, saving costs and time. This work also presents a comparison between numerical and experimental results of acoustic streaming obtained with that ß-PVDF transducer in the movement and in the formation of fluid recirculation in tridimensional closed domains. Differences between the numerical and experimental results are credited to the high sensitivity of acoustic streaming to the experimental conditions and to limitations of the numerical method. The reported study contributes for the improvement of simulation models that can be extremely useful for predicting the acoustic effects of new materials in fluidic devices, as well as for optimizing the transducers and matching layers positioning in a fluidic structure.

Lab-on-a-chip with beta-poly(vinylidene fluoride) based acoustic microagitation.

This paper reports a fully integrated disposable lab-on-a-chip with acoustic microagitation based on a piezoelectric ss-poly(vinylidene fluoride) (ss-PVDF) polymer. The device can be used for the measurement, by optical absorption spectroscopy, of biochemical parameters in physiological fluids. It comprises two dies: the fluidic die that contains the reaction chambers fabricated in SU-8 and the ss-PVDF polymer deposited underneath them; and the detection die that contains the photodetectors, its readout electronics, and the piezoelectric actuation electronics, all fabricated in a CMOS microelectronic process. The microagitation technique improves mixing and shortens reaction time. Further, it generates heating, which also improves the reaction time of the fluids. In this paper, the efficiency of the microagitation system is evaluated as a function of the amplitude and the frequency of the signal actuation. The relative contribution of the generated heating is also discussed. The system is tested for the measurement of the uric acid concentration in urine.

Biological microdevice with fluidic acoustic streaming for measuring uric acid in human saliva.

The healthcare system requires new devices for a rapid monitoring of a patient in order to improve the diagnosis and treatment of various diseases. Accordingly, new biomedical devices are being developed. In this paper, a fully-integrated biological microdevice for uric acid analysis in human saliva is presented. It is based on optical spectrophotometric measurements and incorporates a mixture system based on acoustic streaming, that enhances the fluids reaction due to both heating and agitation generated by this effect. Acoustic streaming is provided by a piezoelectric beta-PVDF film deposited underneath the microfluidic die of the device. Further, it incorporates the electronics for the detection, readout, data processing and signal actuation. Experimental results proved that acoustic streaming based on this piezoelectric polymer is advantageous and reduces in 55% the time required to obtain the analysis results.